Detection and characterization of bovine-like coronaviruses from four species of zoo ruminants

Five coronaviruses (CoVs) were detected in diarrheal feces from four zoo ruminant species: one wisent (Bison bonasus), two Himalayan tahr (Hemitragus jemlahicus), one sitatunga (Tragelaphus spekii), and one nyala (Tragelaphus angasii). We sequenced and analyzed the spike (S) and hemagglutinin/esterase (HE) genes of these viruses and compared the nucleotide (nt) and deduced amino acid (aa) sequences with those of other bovine CoV (BcoV) strains. Comparison of the entire deduced aa sequences of the S and HE glycoproteins revealed no specific differences that would account for discrimination between bovine-like CoV strains from zoo ruminants and BcoVs strains. In addition, the 99.9% aa identity among the five CoV strains revealed that the ruminants were infected by the same strain. Phylogenetically, bovine-like CoVs belong to group 2a CoVs, which are related most closely to the BcoV strains recently isolated in Korea. These data suggest that cattle are potential reservoirs for CoVs that are capable of infecting zoo ruminants.     

One year's study of dynamic and evolution of types I and II PRRSV in a swine farm

This study was to investigate dynamic and evolution of PRRSV in a seed-stock farm by monitoring PRRSV status from 11 June 2009 to 4 August 2010. For laboratory test, around 18-24 umbilical cords from farrowed sows and 5-95 sera from nursery and grow/finish pigs were submitted around every 2 weeks interval during the study. The submitted samples were tested for PRRSV using IDEXX PRRS 2XR ELISA kit, RT-nested PCR. The PRRSV-positive samples were further sequences based on ORF5 and analyzed using MEGA 3.1 program and Beast 1.5.4 package. The surveyed farm was first infected with type II PRRSV but it was infected newly with type I PRRSV of unknown origin, showing rapid substitution to type I PRRSV as a dominant strain in 2 weeks. The type I PRRSV was first detected from umbilical cord of a farrowed sow in 12 January 2010, and secondly from nursery pigs in 26 January 2010. Although sudden increase of mean S/P ratio was found in grow/finish pigs around 2 months earlier than first type I PRRSV detection, no type I PRRSV viremia was found. Thirty three ORF5 full sequences from 14 type II to 19 type I PRRSVs were obtained chronologically in this farm and the genetic characteristics and evolution rates of those sequences were analyzed. The substitution rates (/site/day) of two types were 4.03×10(-5) (type I), 3.09×10(-5) (type II), respectively, which was more frequent than previous reports. The calculated divergence time of type I PRRSV was consistent with the time when the sudden elevation of serum IgG in grow/finish barn was first observed. This study provided fundamental data for type I PRRSV dynamic in a previously type II PRRSV-infected farm and suggested grow/finisher barn could be a primary site for PRRSV introduction.     

Reassortment among bovine, porcine and human rotavirus strains results in G8P[7] and G6P[7] strains isolated from cattle in South Korea

Group A rotaviruses (GARVs) cause severe acute gastroenteritis in children and young animals. Although zoonotic infections with bovine-like G6 and G8 GARVs have been reported in many countries, there is little evidence for reassortment between bovine GARVs and GARVs from heterologous species. The finding of bovine GARVs with the G6 and G8 genotypes in combination with the typical porcine P[7] prompted us to characterize all 11 genes of 30 bovine GARVs isolated from clinically infected calves. By the comparison of the full-length ORF of VP7 and NSP1-5, and the partial VP1-4 and VP6 nucleotide sequences between the 30 Korean and other known strains, three different genome constellations were found. Twenty seven strains showed the G8-P[7]-I5-R1-C1-M2-A1-N1-T1-E1-H1 genotypes, a single strain possessed the G6-P[7]-I2-R2-C1-M2-A1-N2-T1-E2-H1 genotype constellation and 2 strains the G6-P[7]-I2-R2-C2-M2-A3-N2-T6-E2-H3 genotype constellation. The complete genome of a single reference strains for each of these three genotype constellations (KJ25, KJ9-1 and KJ19-2) was determined and analyzed. A detailed phylogenetic analysis revealed a complicated picture, with several reassortments among bovine-like, porcine-like and human-like GARV strains, resulting in several different reassortant strains successfully infecting cattle.     

Genetic diversity of diploid Japanese strawberry species based on microsatellite markers

The United States Department of Agriculture (USDA)—Agricultural Research Service (ARS)—National Clonal Germplasm Repository (NCGR) in Corvallis, Oregon, is a genebank that preserves strawberry genetic resources. Representatives of two Japanese diploid species, Fragaria iinumae Makino and F. nipponica Makino were collected for conservation by the NCGR during an expedition to Hokkaido, Japan. Fragaria iinumae may be a genome contributor to the cultivated octoploid strawberries. The objective of this study was to evaluate the genetic diversity of these two species by using simple sequence repeat (SSR) markers. Twenty of 82 Fragaria-derived SSRs, polymorphic among and within the two species, were selected for genetic analysis of 137 accessions. Genetic diversity, based on the proportion of shared alleles between the two species, in F. nipponica (0.4542) and F. iinumae (0.1808) was significantly different. Three wild interspecific hybrids were identified from intermediate memberships in the two diploid species groups revealed by using the clustering program, Structure. Principal coordinate analysis followed by non-parametric modal clustering (PCO-MC) grouped accessions into two clusters representing the two diploid species. Further clustering within the species groups generated with the program, STRUCTURAMA^TM, resulted in seven subclusters in F. iinumae and three in F. nipponica, which may represent breeding populations appropriate for clonal conservation. Long-term preservation of the species populations and the limited number of hybrids on the island is discussed relative to their geographical distribution and the geological history of Hokkaido Island.     

Insecticidal activity of rhamnolipid isolated from pseudomonas sp. EP-3 against green peach aphid (Myzus persicae)

Microorganisms capable of growth on oils are potential sources of biopesticides, as they produce complex molecules such as biosurfactants and lipopeptides. These molecules have antimicrobial activity against plant pathogens, but few data are available on their insecticidal activity. The present study describes the insecticidal activity of a rhamnolipid isolated from diesel oil-degrading Pseudomonas sp. EP-3 (EP-3). The treatment of cell-free supernatants of EP-3 grown on glucose-mineral medium for 96 h led to > 80% mortality of aphids (Myzus persicae) within 24 h. Bioassay-guided chromatography coupled with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MADLDI-TOF MS) and (1H, 13C) nuclear magnetic resonance (NMR) analyses was employed to isolate and identify the EP-3 insecticidal metabolites. Dirhamnolipid, with molecular formulas of C??H??O?? and C??H??O??, was identified as a main metabolite exhibiting insecticidal activity against aphids. Dirhamnolipid showed a dose-dependent mortality against aphids, producing about 50% mortality at 40 μg/mL and 100% mortality at 100 μg/mL. Microscopy analyses of aphids treated with dirhamnolipid revealed that dirhamnolipid caused insect death by affecting cuticle membranes. This is the first report of rhamnolipid as an insecticidal metabolite against M. persicae. Rhamnolipid shows potential for use as a pesticide to control agricultural pests.     

Modification of curcumin with polyethylene glycol enhances the delivery of curcumin in preadipocytes and its antiadipogenic property

Conjugation of curcumin (CCM) by polyethylene glycol (PEG) has been previously developed to improve water solubility of the natural form of CCM and its antiproliferative role in some human cancer cell lines. This study examined the cellular uptake kinetics of the natural form of CCM and CCM-PEG. Their cytotoxic effect in proliferating preadipocytes and antiadipogenic property in differentiating preadipocytes had also been investigated. CCM and CCM-PEG were found to be differently absorbed in 3T3-L1 preadipocytes and adipocytes with a limited amount of CCM-PEG absorption in the cell. The improved water solubility of CCM-PEG was correlated with increased cellular retention of CCM in 3T3-L1 cells, particularly in preadipocytes. Consequently, CCM-PEG treatment sensitized proliferating preadipocytes to CCM-induced cell toxicity. Furthermore, incubation of differentiating 3T3-L1 cells with CCM-PEG resulted in improvement of the inhibitory role of CCM in adipocyte differentiation with no toxic effect. These results suggest that pegylation-improved water solubility and cellular retention of CCM may be uniquely useful for improving the delivery of CCM in preadipocytes and its antiadipogenic ability.     

The impact of packaging materials on the antioxidant phytochemical stability of aqueous infusions of green tea (Camellia sinensis) and yaupon holly (Ilex vomitoria) during cold storage

Ready to drink (RTD) teas are a growing segment in the beverage category, brought about by improvements in the flavor of these products and healthy market trends driven by consumers. The presented results evaluated the antioxidant phytochemical stability of RTD teas from aqueous infusions of traditional green tea (Camellia sinensis) and a botanical tea from yaupon holly (Ilex vomitoria) as influenced by packaging materials during cold storage. Two common packaging materials for RTD products are glass and polyethylene terephthalate (PET) and have been compared to a retortable pouch (RP), an emerging packaging material for various types of food since it is durable, inexpensive, lightweight, and easy to sterilize. Storage stability was then evaluated for each aqueous infusion prepared at 10 g/L at 90 °C for 10 min and evaluated at 3 °C in the absence of light over 12 weeks. Analyses included quantification and characterization of individual polyphenolics by high-performance liquid chromatography-photodiode array and liquid chromatography-electrospray ionization-mass spectrometry as well as changes in total antioxidant capacity. For green tea, concentrations of the three major flavan-3-ols, epigallocatechin gallate, epigallocatechin, and epicatechin gallate were better retained in glass bottles as compared to other packages over 12 weeks. In yaupon holly, chlorogenic acid and its isomers that were the predominant compounds were generally stable in each packaging material, and a 20.6-fold higher amount of saponin was found as compared to green tea, which caused higher stability of flavonol glycosides present in yaupon holly during storage. The antioxidant capacity of green tea was better retained in glass and PET versus RP, whereas no differences were again observed for yaupon holly. Results highlight the superiority of oxygen-impervious glass packaging, but viable alternatives may be utilizable for RTD teas with variable phytochemical compositions.     

Occurrence and Environmental Fate of Veterinary Antibiotics in the Terrestrial Environment

A wide variety of veterinary antibiotics (VAs) has been detected in environmental water samples, and this is of potential environmental concern due to their adverse effects. In particular, the potential for development of antibiotic-resistant bacteria has raised social concerns leading to intensive investigation regarding the influence of antibiotics on human and ecosystem health. One of the main sources of antibiotic effluence to the environment is livestock manures that often contain elevated levels of VAs that survive normal digestive procedures following medication in animal husbandry because unlike human waste, waste generated on farms does not undergo tertiary wastewater treatment, and consequently, the concentration of antibiotics entering the environment is expected to be larger from farming practices. Animal feed is often supplemented with VAs to promote growth and parasite resistance in the medicated animals, and this practice typically resulted in higher use of VAs and consequential excretion from livestock through urine and feces. The excretion rate varied depending on the type of VA used with around 75, 90, and 50?C100% being excreted for chlortetracycline, sulfamethazine, and tyolsin, respectively. The excreted VAs that initially present in livestock manures were degraded more than 90% when proper composting practice was used, and hence, this can be employed as a management strategy to decrease VA environmental loads. The reduction of VA concentrations during composting was mainly attributed to abiotic processes rather than biotic degradation. The VAs released to soils by the application of manure and manure-based composts can be degraded or inactivated to various degrees through abiotic process such as adsorption to soil components. Depending on the antibiotic species and soil properties, residues can be transferred to groundwater and surface water through leaching and runoff and can potentially be taken up by plants.     

Removal of Phosphate from Aqueous Solution by Functionalized Mesoporous Materials

In the present study, the applications of mesoporous materials based on silica, namely post-synthesized, one-pot synthesized, and pure MCM-41, were investigated for the removal of phosphate from aqueous solution. The mesostructures were confirmed by X-ray diffraction, Brunauer?CEmmett?CTeller, Fourier transform spectroscopy, and transmission electron microscopy. The absorptions of phosphate by the mesoporous adsorbents were examined, with different adsorption models used to describe the equilibrium and kinetic data. The maximum adsorption capacities of the mesostructure adsorbents were found to be 45.162, 40.806, and 31.123 mg g^?C1 for the post-synthesized, one-pot synthesized, and pure MCM-41, respectively. The kinetic data showed that the adsorptions of phosphate onto the post-synthesized and pure MCM-41 followed the pseudo-first-order kinetic model, whereas the one-pot synthesized adsorbent was described by the pseudo-second-order model.